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To identify the molecular basis of its lipid-binding function, we determined the first crystal structure at 2.49\u2005\u00c5 resolution for fatty acid-bound ZAG, where the ligand was the fluorescent 11-(dansylamino)undecanoic acid (DAUDA). The 192\u2005kDa crystallographic asymmetric unit contained six ZAG and eight fatty acid molecules in unique conformations. Six fatty acid molecules were localised to the ZAG grooves, where their tails were bound in two distinct conformations. The carboxylate groups of three fatty acids projected out of the groove, while the fourth was hydrogen bonded with R73 inside the groove. Other ligand-residue contacts were primarily hydrophobic. A new fatty acid site was revealed for two further DAUDA molecules at the ZAG \u03b13 domains. Following conformational changes from unbound ZAG, the \u03b13 domains formed tetrameric \u03b2-barrel structures lined by fatty acid molecules that doubled the binding capacity of ZAG. Analytical ultracentrifugation revealed that ZAG in solution was a monomer in the absence of DAUDA, but formed small amounts of tetramers with DAUDA. By showing that ZAG binds fatty acids in different locations, we demonstrate an augmented mechanism for fatty acid binding in ZAG that is distinct from other known fatty acid binding proteins, and may be relevant to cachexia.<\/jats:p>","DOI":"10.1042\/bcj20190354","type":"journal-article","created":{"date-parts":[[2019,9,10]],"date-time":"2019-09-10T16:05:21Z","timestamp":1568131521000},"page":"2815-2834","update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Crystal structure of zinc-\u03b12-glycoprotein in complex with a fatty acid reveals multiple different modes of protein-lipid binding"],"prefix":"10.1042","volume":"476","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0572-7826","authenticated-orcid":false,"given":"Andy M.","family":"Lau","sequence":"first","affiliation":[{"name":"Department of Structural and Molecular Biology, University College London, Darwin Building, Gower Street, London WC1E 6BT, U.K."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6882-3335","authenticated-orcid":false,"given":"Henna","family":"Zahid","sequence":"additional","affiliation":[{"name":"Department of Structural and Molecular Biology, University College London, Darwin Building, Gower Street, London WC1E 6BT, U.K."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1654-1392","authenticated-orcid":false,"given":"Jayesh","family":"Gor","sequence":"additional","affiliation":[{"name":"Department of Structural and Molecular Biology, University College London, Darwin Building, Gower Street, London WC1E 6BT, U.K."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9218-9805","authenticated-orcid":false,"given":"Stephen J.","family":"Perkins","sequence":"additional","affiliation":[{"name":"Department of Structural and Molecular Biology, University College London, Darwin Building, Gower Street, London WC1E 6BT, U.K."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7385-0143","authenticated-orcid":false,"given":"Alun R.","family":"Coker","sequence":"additional","affiliation":[{"name":"Division of Medicine, University College London, The Rayne Building, 5 University Street, London WC1E 6JF, U.K."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4723-6056","authenticated-orcid":false,"given":"Lindsay C.","family":"McDermott","sequence":"additional","affiliation":[{"name":"School of Life Sciences, University of Bedfordshire, Park Square, Luton LU1 3JU, U.K."}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"288","published-online":{"date-parts":[[2019,10,11]]},"reference":[{"key":"2021112218005499700_BCJ-2019-0354C1","doi-asserted-by":"crossref","first-page":"1066","DOI":"10.1016\/S0021-9258(18)64243-7","article-title":"Preparation and properties of Zn-\u03b12-glycoprotein of normal human plasma","volume":"236","year":"1961","journal-title":"J. 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